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4. 4a shows the DIHM reconstruction procedure, from raw data (on the left) through to the ﬁtted contour that represents the shape of the microgamete (on the right-hand side). Assuming that the microtubules are bound together at one end (the ‘no basal sliding’ approximation), the relative displacement of microtubules within the microgamete can be extracted from the flagellar shape, as seen in Fig. 4b. The colour map shows the sliding of microtubule doublets in the axoneme relative to the gamete centre line, from the relatively passive ‘tail’ end of the microgamete (position s ¼ 0 µm on the horizontal axis) to the more active head end (s ¼ 9 µm).

2006) found a MIC of 4 μM for pexiganan acting on E. 67 μM after only 5 min of incubation. Ramamoorthy et al. (2006) infer that factors other than membrane disruption must be involved in the bacterial killing process. A later study came to a similar conclusion on similar grounds (Pius et al. 2012). 2 × MIC. In general, understanding what MIC values mean is crucial if pharmacodynamic results are to be correctly compared with mechanistic studies. Thirdly, there is strong evidence that peptides causing cell death are sequestered and unavailable for further bactericidal action.

3 Data Processing Raw DIHM images are two-dimensional and look similar to out-of-focus bright ﬁeld microscope images (see Fig. 1). These images contain information about the three-dimensional positions and shapes of the cells in the sample chamber. There are several computational processing schemes for extracting three-dimensional information from holographic images (Lee et al. 2007; Kim 2010), but we have found the Rayleigh-Sommerfeld back-propagation method (Lee and Grier 2007) to be the most convenient.